PMID- 27327271
OWN - NLM
STAT- MEDLINE
DCOM- 20170719
LR  - 20190213
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 11
IP  - 6
DP  - 2016
TI  - Redesigning Aldolase Stereoselectivity by Homologous Grafting.
PG  - e0156525
LID - 10.1371/journal.pone.0156525 [doi]
LID - e0156525
AB  - The 2-deoxy-d-ribose-5-phosphate aldolase (DERA) offers access to highly 
      desirable building blocks for organic synthesis by catalyzing a stereoselective 
      C-C bond formation between acetaldehyde and certain electrophilic aldehydes. 
      DERA s potential is particularly highlighted by the ability to catalyze 
      sequential, highly enantioselective aldol reactions. However, its synthetic use 
      is limited by the absence of an enantiocomplementary enzyme. Here, we introduce 
      the concept of homologous grafting to identify stereoselectivity-determining 
      amino acid positions in DERA. We identified such positions by structural analysis 
      of the homologous aldolases 2-keto-3-deoxy-6-phosphogluconate aldolase (KDPG) and 
      the enantiocomplementary enzyme 2-keto-3-deoxy-6-phosphogalactonate aldolase 
      (KDPGal). Mutation of these positions led to a slightly inversed 
      enantiopreference of both aldolases to the same extent. By transferring these 
      sequence motifs onto DERA we achieved the intended change in enantioselectivity.
FAU - Bisterfeld, Carolin
AU  - Bisterfeld C
AD  - Institut fur Bioorganische Chemie, Heinrich-Heine-Universitat Dusseldorf im 
      Forschungszentrum Julich, 52426, Julich, Germany.
FAU - Classen, Thomas
AU  - Classen T
AD  - Institute of Bio- and Geosciences IBG-1: Biotechnology, Forschungszentrum Julich 
      GmbH, 52425, Julich, Germany.
FAU - Kuberl, Irene
AU  - Kuberl I
AD  - Institut fur Bioorganische Chemie, Heinrich-Heine-Universitat Dusseldorf im 
      Forschungszentrum Julich, 52426, Julich, Germany.
FAU - Henssen, Birgit
AU  - Henssen B
AD  - Institut fur Bioorganische Chemie, Heinrich-Heine-Universitat Dusseldorf im 
      Forschungszentrum Julich, 52426, Julich, Germany.
FAU - Metz, Alexander
AU  - Metz A
AD  - Institute of Pharmaceutical and Medicinal Chemistry, Heinrich-Heine-Universitat 
      Dusseldorf, Dusseldorf, Germany.
FAU - Gohlke, Holger
AU  - Gohlke H
AD  - Institute of Pharmaceutical and Medicinal Chemistry, Heinrich-Heine-Universitat 
      Dusseldorf, Dusseldorf, Germany.
FAU - Pietruszka, Jorg
AU  - Pietruszka J
AD  - Institut fur Bioorganische Chemie, Heinrich-Heine-Universitat Dusseldorf im 
      Forschungszentrum Julich, 52426, Julich, Germany.
AD  - Institute of Bio- and Geosciences IBG-1: Biotechnology, Forschungszentrum Julich 
      GmbH, 52425, Julich, Germany.
LA  - eng
PT  - Journal Article
DEP - 20160621
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Amino Acids)
RN  - 0 (Pyruvates)
RN  - EC 4.1.2.- (Aldehyde-Lyases)
RN  - EC 4.1.2.13 (Fructose-Bisphosphate Aldolase)
RN  - EC 4.1.2.14 (phospho-2-keto-3-deoxy-gluconate aldolase)
SB  - IM
MH  - Aldehyde-Lyases/chemistry/metabolism
MH  - Amino Acid Sequence
MH  - Amino Acids/metabolism
MH  - Biocatalysis
MH  - Escherichia coli/enzymology
MH  - Fructose-Bisphosphate Aldolase/chemistry/*metabolism
MH  - Kinetics
MH  - Models, Molecular
MH  - Phylogeny
MH  - *Protein Engineering
MH  - Protein Structure, Secondary
MH  - Pyruvates/metabolism
MH  - Stereoisomerism
MH  - Substrate Specificity
PMC - PMC4915726
COIS- Competing Interests: The authors have declared that no competing interests exist.
EDAT- 2016/06/22 06:00
MHDA- 2017/07/20 06:00
PMCR- 2016/06/21
CRDT- 2016/06/22 06:00
PHST- 2016/02/08 00:00 [received]
PHST- 2016/05/16 00:00 [accepted]
PHST- 2016/06/22 06:00 [entrez]
PHST- 2016/06/22 06:00 [pubmed]
PHST- 2017/07/20 06:00 [medline]
PHST- 2016/06/21 00:00 [pmc-release]
AID - PONE-D-16-05568 [pii]
AID - 10.1371/journal.pone.0156525 [doi]
PST - epublish
SO  - PLoS One. 2016 Jun 21;11(6):e0156525. doi: 10.1371/journal.pone.0156525. 
      eCollection 2016.